[Investigate the role of neutrophil extracellular traps in immune checkpoint inhibitor-associated myocarditis with programmed death protein-1 inhibitors involvement].

Objective: To investigate the role of neutrophil extracellular traps (NETs) in immune checkpoint inhibitor-associated myocarditis (ICIAM) with programmed death protein-1 (PD-1) inhibitors involvement, and to explore the therapeutic potential of targeting NETs in the treatment of ICIAM. Methods: Thirty 6-week-old male BALB/c mice were randomly divided into control group (n=10), myocarditis group (n=10), and treatment group (n=10). Apart from the control group, each mouse was subcutaneously injected with 100 µl of complete Freund's adjuvant containing 250 µg of mouse cardiac troponin I peptide on the 1st and 7th day. Starting on the 8th day, PD-1 inhibitor (15 µg/per mouse) was intraperitoneally injected every other day for a total of 5 times. Since 1 day before the beginning of PD-1+TnI injection, the treatment group was injected with PF-1355 (50 mg·kg-1·d-1) for 16 consecutive days. The mice's general state was observed during the whole process. Real-time fluorescence quantitative PCR (RTFQ-PCR) was carried out to evaluate the transcriptional regulation of neutrophil related chemokines, NETs, pyronecrosis related factors and proinflammatory cytokines. Immunohistochemistry, immunofluorescence and western blot were applied to determine the changes of pyrosis related molecules. Echocardiography showed the differences of main cardiac indexes while cardiac pathology compared the degree of inflammatory infiltration in 3 gruops. Results: The immunofluorescence intensity of myocardial NETs in the myocarditis group was significantly increased compared to the control group mice (2.49±0.08 and 0.99±0.26, P<0.001). The protein expression levels of pyroptosis-related NLRP3, cleaved-Caspase 1, Caspase 1, cleaved-GSDMD, GSDMD, IL-1ß and IL-18 in myocardial tissue of the model group were higher than those of the control group (all P<0.05). After treatment with PF-1355, compared to the myocarditis group, the left ventricular ejection fraction (LVEF) (73.58%±5.31% and 58.12%±3.19%, P<0.001) and left ventricular fraction shortening (LVFS) (39.78%±4.31% and 33.89%±2.19%, P<0.001) increased. H-E staining showed a reduction in inflammatory infiltration area in the treatment group compared to the myocarditis group (30.12%±3.57% and 14.92%±2.46%, P<0.001). The immunofluorescence intensity of NETs decreased in the treatment group compared to the myocarditis group (2.52±0.04 and 1.03±0.05, P<0.001). The levels of NLRP3 and other pyroptosis-related molecules were downregulated in the treatment group compared to the myocarditis group (all P<0.05). Conclusions: NETs lead to myocardial cell pyroptosis by activating the NLRP3 inflammasome in PD-1 inhibitor-associated myocarditis. The specific MPO inhibitor PF-1355 shows a therapeutic potential by regulating the formation of NETs, decreasing NLRP3 level and relieving myocardial pyroptosis, thus reducing myocardial damage.

[Preliminary practice in the postoperative cardiopulmonary exercise testing for children with congenital heart disease].

Objective: To investigate the postoperative cardiopulmonary fitness of children with congenital heart diseases (CHD). Methods: This is a retrospective study. A total of 136 children after cardiac correction undergoing cardiopulmonary exercise test (CPET) in Shanghai Children's Medical Center from March 1 to June 30, 2021 were selected. According to the surgical procedure, the children were divided into two groups: the biventricular correction group (BV group) (n=75) and single ventricular correction group (SV group) (n=61). The BV group was divided into two subgroups: simple congenital heart disease (S-CHD) group (n=35) and complex congenital heart disease (C-CHD) group (n=40). CPET parameters, including VO2 max, VO2/kg max, VO2/kg@AT, O2/HR max, HRR, PetCO2 max, CI, HRR at 1 min, VE/VCO2 slope, OUES/kg and EOV, were analyzed. CPET parameters of patients underwent different procedure were compared. In the subgroup analysis, CPET parameters of patients in S-CHD group and C-CHD group were compared. The parameters of the BV group were compared with the normal value. Linear correlation analysis was used to identify the correlation between the CPET parameters. Results: A total of 136 children with CHD after surgery were enrolled. The age was (9.4±3.1) (ranged 6.2-16.0) years, and there was 84 (61.8%) male. All the children completed CPET examination safely without experiencing serious circulation abnormalities. Compared with the SV group, the VO2/kg max ((32.6±6.9) ml·kg-1·min-1 vs. (23.5±5.9) ml·kg-1·min-1, P<0.001), O2/HR max ((7.24±2.93) ml/beat vs. (6.35±2.17)ml/beat, P=0.030), HRR at 1 min ((32.5±13.9) beat/min vs. (26.3±12.5) beat/min, P=0.036), OUES/kg (36.9±8.8 vs. 29.7±11.8, P=0.001) were significantly higher, VE/VCO2 slope (29.1±5.20 vs. 35.1±8.0, P<0.001) and incidence of EOV (32.0%(24/75) vs. 57.4%(31/61), P=0.027) were significantly lower in BV group. Compared with the simple CHD subgroup, VE/VCO2 slope and the incidence of EOV were higher, VO2/kg max, O2/HR max, HRR at 1 min and OUES/kg were lower in the complex CHD subgroup (all P<0.05). Cardiopulmonary function parameters of the BV group were lower than the normal value. Linear correlation analysis showed that VO2/kg@AT was strongly correlated with VO2/kg max (r=0.86, P<0.001), VO2/kg max was strongly correlated with OUES/kg (r=0.63, P<0.001), HRR was strongly correlated with CI (r=0.91, P<0.001), and VO2/kg max was strongly correlated with OUES/kg (r=0.63, P<0.001). VE/VCO2 slope was strongly correlated with PetCO2 max (r=1.00, P<0.001). Conclusions: The exercise cardiopulmonary function of children after single ventricular correction is weaker than that of biventricular correction, and the exercise tolerance of children after biventricular correction is lower than that of normal children. Among the children after biventricular correction, the exercise tolerance of children with complex CHD is lower than that of children with simple CHD. Postoperative CPET is of important realistic significance for CHD children.

[Analysis of lactate dehydrogenase gene polymorphisms and prediction of B cell epitopes in four human Plasmodium species].

OBJECTIVE: To analyze the polymorphism of Plasmodium lactate dehydrogenase (pLDH) gene and predict B-cell epitopes in pLDH peptides in four species of human malaria parasites. METHODS: The blood samples and epidemiological characteristics were collected from malaria cases in Yunnan Province registered in the National Notifiable Disease Report System. The pLDH genes of four human Plasmodium species were amplified using nested PCR assay and sequenced. The polymorphisms of pLDH genes was analyzed using the software MEGA version 7.0.26 and DnaSP version 5.10, and the B-cell epitopes were predicted in pLDH peptides using the Immune Epitope Database (IEDB). RESULTS: The sequences of P. vivax LDH (PvLDH), P. falciparum LDH (PfLDH), P. ovale LDH (PoLDH) and P. malariae LDH (PmLDH) genes were obtained from 153, 29, 17 and 11 blood samples from patients with P. vivax, P. falciparum, P. ovale and P. malariae malaria, respectively, which included 15, 2, 4 and 2 haplotypes and had a nucleotide diversity (π) of 0.104. A high level of intra-species differentiation was seen in the PoLDH gene (π = 0.012), and the π values were all < 0.001 for PvLDH, PfLDH and PmLDH genes. Active regions of B-cell antigen were predicted in the pLDH peptide chain of four human malaria parasites, of 4 to 5 in each chain, and the activity score was approximately 0.430. Among these peptide chains, the "86-PGKSDKEWNRD-96" short-peptide was a B-cell epitope shared by all four species of human malaria parasites, and the "266-GQYGHS (T)-271" short-peptide was present in PvLDH and PoLDH peptide chains, while "212-EEVEGIFDR-220" was only found in the PvLDH peptide chain, and "208-LISDAE-213" was only seen in the PfLDH peptide chain. CONCLUSIONS: The PoLDH gene polymorphism may be derived from the weak negative purification selection, while PvLDH, PfLDH and PmLDH genes may maintain a relatively conservative state. There may be two B-cell epitopes "212-EEVEGIFDR-220" and "208-LISDAE-213" in the proximal region of the C terminal in the pLDH peptide chain, which is feasible to differentiate between P. vivax and P. falciparum infections.

[Genomic investigation of human Streptococcus suis infection in Shandong Province].

To investigate Streptococcus suis (S.suis) isolated from patients in Shandong province using genomic epidemiology and pathogenologic analysis. To provide the foundation to establish reasonable and accurate prevention and control measures of human S. suis infection. Molecular typing, whole genome phylogenetic tree, virulence gene typing, antibiotic resistance profile and mobile genetic elements carrying antibiotic resistance genes of isolated S. suis strains were investigated. The pathogenicity of isolated strains was also evaluated by comparing their capacity to induce pro-inflammatory cytokine production in vitro. S. suis infections in Shandong province were predominantly due to serotype 2 and sequence type 1 strains. The major symptoms were meningitis. The studied strains could be divided into five lineages. All strains belong to highly pathogenic type in Shandong province,Strains from lineage 2 possessed higher capacity to stimulate pro-inflammatory cytokine production than other strains did, even though other strains belong to highly pathogenic strains. In addition, multiple antibiotic resistance genes and corresponding mobile genetic elements werewidespread in S. suis strains from Shandong province, except strains from lineage 3. High diversities in genome, evolutionary path and pathogenicity of S. suis strains from Shandong province were revealed. It was necessary to surveillant the S. suis strain in genomic level.

[Interpretation of group standard for Clostridioides difficile infection diagnosis].

Clostridioides difficile is a key pathogen of antibiotic related diarrhea and hospital associated infection, causing several outbreaks in Europe and North Americans and resulting in severe disease burden. However, the standardized diagnostic principle and detection specifications in C. difficile infection (CDI) survey are limited in China, and the infection rate and disease burden of CDI in China are unclear. Therefore, National Institute for Communicable Disease Control and Prevention,National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, together with another 11 institutions, draft the group standard entitled "Diagnosis of Clostridium difficile infection (T/CPMA 008-2020)" of Chinese Preventive Medicine Association. Based on the principle of "legality, scientificity, advancement, and feasibility", this standard clarifies risk factors, diagnosis principles, diagnoses and differential diagnoses in order to improve the accuracy of CDI diagnosis in clinical practice, guide the surveillance for CDI, and understand the infection rate and disease burden of CDI in China.

[Etiological survey and traceability analysis of five cases of typhoid fever during the same period in Yantai city].

Objective: To analyze the etiological characteristics and molecular epidemiological correlation of five cases of typhoid fever during the same period in yantai city. Methods: Six S. Typhis strains were isolated from 5 typhoid patients and epidemiological samples in Yantai city in 2018. The onset time of the cases were from May 26, 2018 to July 24, 2018, distributed in Shuidao Town of Muping District, Dengzhou Street of Penglai District, Donglai Street of Longkou District, Wenhua Street of Muping District and Fulaishan Street of zhifu District. S. Typhis strains were analyzed by conventional bacterial isolation method and XbaⅠ/BlnⅠ double-enzyme digestion pulse-field gel electrophoresis (PFGE). Meanwhile, ViaB virulence gene detection and 27 common antibiotics sensitivity tests were conducted to study the etiology of S. Typhis. Results: Six strains of S. Typhi were isolated from 5 patients and the domestic egg of one patient, which were divided into 4 PFGE patterns by PFGE-XbaⅠ and PFGE-BlnⅠ and among which 3 strains had the same PFGE patterns.One multi-drug resistant strain (foreign patient), one single-drug resistant strain (patient with a history of provincial retention), and one completely sensitive strain were detected. The three strains of the same PFGE pattern exhibit the same drug-sensitive phenotype which were intermediate against aminoglycosides and quinolones and susceptibility against the other antibiotics.All of the strains carried the ViaB virulence factor except the strain from the foreign patient. Conclusion: Local S. Typhi is susceptibility or intermediate against antibiotics commonly used in clinic.Sporadic cases of typhoid fever and typhoid imported infections still need attention.

Neuroprotective effect of CTRP3 overexpression against sevoflurane anesthesia-induced cognitive dysfunction in aged rats through activating AMPK/SIRT1 and PI3K/AKT signaling pathways.

OBJECTIVE: To investigate the effects of C1q/tumor necrosis factor-related protein-3 (CTRP3) on postoperative cognitive dysfunction (POCD) and elucidate the potential regulatory mechanism in sevoflurane anesthesia-induced aged rats. MATERIALS AND METHODS: A sevoflurane anesthesia-induced POCD aged rat model was established and hematoxylin and eosin (H&E) staining was used to detect pathological changes of hippocampal neurons. Morris water maze task test was performed to determine the learning and memory ability of rats. Immunofluorescence, quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) and Western blot were used to detect CTRP3 expression. Enzyme-linked immunosorbent assay (ELISA) or qRT-PCR assays were used to evaluate the changes of markers of brain damage and inflammatory cytokines. Terminal deoxynucleotidyltransferase-mediated dUTP nick-end labeling (TUNEL) assay was used to assess the apoptosis of nerve cells in hippocampus. Western blot assay were used to measure the expression levels of apoptosis-related protein, and AMP-activated protein kinase (AMPK)/SIRT1 and PI3K/AKT pathway. RESULTS: Sevoflurane exposure led to brain injury, cognitive dysfunction in aged rats and decreased the expression of CTRP3. Overexpression of CTRP3 could suppress nerve cell apoptosis, inhibit neuronal inflammation, reduce brain tissue damage and improve cognitive dysfunction of aged rats after sevoflurane anesthesia. Further studies showed that CTRP3 may play a role in POCD by regulating AMPK/SIRT1 and PI3K/AKT signaling pathways. CONCLUSIONS: CTRP3 may effectively protect against sevoflurane-induced cognitive dysfunction and served as a potential predictive indicator and therapy target for POCD.

Adjuvant chemotherapy for patients with gastric neuroendocrine carcinomas or mixed adenoneuroendocrine carcinomas.

BACKGROUND: The aim of this study was to evaluate whether adjuvant chemotherapy is associated with improved survival in patients with resectable gastric neuroendocrine carcinomas (G-NECs) or mixed adenoneuroendocrine carcinomas (G-MANECs). METHODS: The study included patients with G-NECs or G-MANECs who underwent surgery in one of 21 centres in China between 2004 and 2016. Propensity score matching analysis was used to reduce selection bias, and overall survival (OS) in different treatment groups was estimated by the Kaplan-Meier method. RESULTS: In total, 804 patients with resectable G-NECs or G-MANECs were included, of whom 490 (60·9 per cent) received adjuvant chemotherapy. After propensity score matching, OS in the chemotherapy group was similar to that in the no-chemotherapy group. Among patients with G-NECs, survival in the fluorouracil (5-FU)-based chemotherapy group and the non-5-FU-based chemotherapy group was similar to that in the no-chemotherapy group. Similarly, etoposide plus cisplatin or irinotecan plus cisplatin was not associated with better OS in patients with G-NECs. Among patients with G-MANECs, OS in the non-5-FU-based chemotherapy group was worse than that in the no-chemotherapy group. Patients with G-MANECs did not have better OS when platinum-based chemotherapy was used. CONCLUSION: There was no survival benefit in patients who received adjuvant chemotherapy for G-NECs or G-MANECs.

ANTECEDENTES: El objetivo de este estudio fue evaluar si la quimioterapia adyuvante mejoraba la supervivencia en pacientes con carcinomas gástricos resecables neuroendocrinos (gastric neuroendocrine carcinomas, G-NECs) y carcinomas adenoneuroendocrinos mixtos (mixed adenoneuroendocrine carcinomas, G-MANECs). MÉTODOS: Se incluyeron pacientes con G-NECs y G-MANECs tratados quirúrgicamente en 21 centros en China entre 2004 y 2016. Se utilizó un análisis de emparejamiento por puntaje de propensión para reducir el sesgo de selección y el método de Kaplan-Meier para estimar la supervivencia global (overall survival, OS) de los pacientes en los diferentes grupos de tratamiento. RESULTADOS: En total, se incluyeron en el estudio 804 pacientes con G-NECs y G-MANECs resecables y 490 pacientes (60,9%) recibieron quimioterapia adyuvante. Después del emparejamiento por puntaje de propensión, la OS del grupo con quimioterapia fue similar a la del grupo sin quimioterapia. En los pacientes con G-NECs, la supervivencia en los grupos con quimioterapia basada en 5-FU (fluorouracilo) y de quimioterapia sin 5-FU fue similar a la del grupo sin quimioterapia. Asimismo, la combinación de etopósido y cisplatino o de irinotecán y cisplatino no se asoció con una mejor OS en pacientes con G-NECs. En pacientes con G-MANECs, la OS del grupo con quimioterapia sin 5-FU fue peor que la del grupo sin quimioterapia. Los pacientes con G-MANECs no presentaron una mejor OS cuando se administró quimioterapia basada en platinos. CONCLUSIÓN: La administración de quimioterapia adyuvante en pacientes con G-NECs y G-MANECs no mejoró la supervivencia.

Long noncoding RNA NEAT1 functions as an oncogene in human laryngocarcinoma by targeting miR-29a-3p.

OBJECTIVE: Long noncoding RNAs (lncRNAs) have been reported to participate in the progression and development of many human diseases. In this study, we are committed to uncover the potential function of lncRNA Nuclear Enriched Abundant Transcript 1 (NEAT1) in the development of laryngocarcinoma. PATIENTS AND METHODS: LncRNA NEAT1 expression in laryngocarcinoma cells and 54 paired laryngocarcinoma samples was detected by Real-time quantitative polymerase chain reaction (RT-qPCR). Furthermore, the regulatory effects of NEAT1 on the proliferation and metastasis of laryngocarcinoma cells were evaluated. Biological role of NEAT1/miR-29a-3p axis was finally explored in regulating the progression of laryngocarcinoma. RESULTS: NEAT1 was upregulated in laryngocarcinoma tissues and cell lines. NEAT1 knockdown suppressed growth and invasive abilities in laryngocarcinoma cells, while overexpression of NEAT1 enhanced such abilities. Further experiments showed that miR-29a-3p was directly targeted by NEAT1, and participated in NEAT-mediated progression of laryngocarcinoma. CONCLUSIONS: NEAT1 is a novel oncogene in laryngocarcinoma and could enhance growth and invasion of laryngocarcinoma cells by targeting miR-29a-3p.

LncRNA SNHG1 promotes cell proliferation in laryngeal cancer via Notch1 signaling pathway.

OBJECTIVE: We aimed at elucidating the potential function of long noncoding ribonucleic acids (lncRNAs) small nucleolar RNA host gene 1 (SNHG1) in the progression of laryngeal cancer (LC) and its underlying mechanism. PATIENTS AND METHODS: Relative level of SNHG1 in LC tissues and controls was determined by quantitative real-time polymerase chain reaction (qRT-PCR). Its expression in LC patients with different tumor stages and statues of lymph node metastasis was examined as well. Correlation between SNHG1 expression and prognosis of LC patients was evaluated by the Kaplan-Meier method. SNHG1 siRNA (si-SNHG1) was constructed for downregulation of SNHG1 expression. Potential effects of downregulated SNHG1 on viability and proliferation of LC cells were detected by cell counting kit-8 (CCK-8) and colony formation assay, respectively. After knockdown of SNHG1, relative levels of Notch1 and hairy, and enhancer of split homolog-1 (Hes1) were determined by qRT-PCR and Western blot. Regulatory effects of SNHG1/Notch1 axis on biological behaviors of LC were finally evaluated. RESULTS: SNHG1 was upregulated in LC tissues than that of controls. Besides, its level was higher in LC with T3-T4 relative to those of T1-T2. Higher abundance of SNHG1 was identified in LC patients with lymph node metastasis compared with those non-metastatic patients. Survival analysis indicated that LC patients with high-level SNHG1 had worse overall survival. Knockdown of SNHG1 in Tu212 and Hep2 cells downregulated relative levels of Notch1 and Hes1. Moreover, SNHG1 knockdown resulted in decreased viability and proliferative ability of LC cells. Notch1 overexpression could reverse the regulatory effects of SNHG1 on viability and proliferation of LC cells. CONCLUSIONS: LncRNA SNHG1 is highly expressed in LC tissues. It promotes the proliferation of LC cells by inhibiting Notch1 pathway, thereby promoting the progression of LC.

[Genetic characterization of influenza A virus and assessment of vaccine efficacy in Yantai from 2014 to 2017].

In this study, the swabs were collected among patients with an influenza-like illness (ILI) admitted to 2 sentinel surveillance hospitals of Yantai from April 2014 to August 2017. All specimen were cultured and identified by hemagglutination inhibition assay. Complete sequences of Hemagglutinin (HA) of influenza A were amplified, sequenced and analyzed using molecular and phylogenetic methods. The potential vaccine efficacy were calculated using Pepitope model. The results showed that the antigenicity of A (H3N2) had changed greatly. 8 strains of influenza A (H1N1) pdm09 belonged to subclade 6B.1 and 14 strains clustered in 6B.2. 12 strains of influenza A (H3N2) fell into subgroup 3C.3a and 33 strains clustered in 3C.2a. Several residues at antigen sites and potential glycosylation sites had changed in influenza A strains. Vaccine efficacy of influenza A (H1N1) pdm09 in 2015/2016 and 2016/2017 seasons were 77.29% and 79.11% of that of a perfect match with vaccine strain, meanwhile vaccine efficacy of influenza A (H3N2) in 2014/2015, 2015/2016 and 2016/2017 were-5.18%, 16.97% and 42.05% separately. In conclusion, the influenza A virus circulated in Yantai from 2014 to 2017 presented continual genetic variation. The recommended vaccine strains still afforded protection against influenza A (H1N1) pdm09 strains and provided suboptimal protection against influenza A (H3N2) strains.

Detection of bone marrow oedema in knee joints using a dual-energy CT virtual non-calcium technique.

AIM: To evaluate bone marrow oedema in knee joints quantitatively and qualitatively using a dual-energy computed tomography (CT) virtual non-calcium (VNCa) technique. MATERIALS AND METHODS: Thirty-five patients with knee joint injuries underwent both dual-energy CT and magnetic resonance imaging (MRI) between March 2018 and November 2018. The presence of bone marrow oedema was assessed by two independent radiologists with the use of colour-coded dual-energy CT VNCa images and measured attenuation on them. The biggest area of bone marrow oedema on axial images was measured by another radiologist using the dual-energy CT and MRI images, respectively. Attenuation values were subjected to receiver operating characteristic (ROC) curve analysis and oedema area sizes were subjected to paired t-test analysis. RESULTS: In qualitative analysis, colour-coded VNCa images had an overall sensitivity of 88.4%, specificity of 98%, positive predictive value of 92.7%, negative predictive value of 96.8%, and accuracy of 95.9%. Attenuation values obtained from colour-coded VNCa images were significantly different in knee joint regions with and without oedema (p<0.001). ROC curve analysis revealed an area under the curve (AUC) of 0.910. A cut-off value of -67 HU provided a sensitivity of 81.4%, specificity of 99.3%, accuracy of 90.4%, positive predictive value of 99.1%, and negative predictive value of 84.2% for the differentiation of oedematous knee joint regions. Significant differences in the size of oedema area were not found between them. CONCLUSION: Dual-energy CT VNCa can be used to evaluate bone marrow oedema effectively.

FBLN3 inhibited the invasion and metastasis of colorectal cancer through the AKT/mTOR pathway.

Fibulin-3(FBLN3) levels are different in different types of cancers. We found that fibulin-3 was downregulated in colorectal (CRC) cells, particularly in the SW480 cell line. By comparison, transfecting SW480 cells with a lentivirus overexpressing fibulin-3 RNA could inhibit proliferation, induce G1/S arrest, and promote cell apoptosis. Fibulin-3 overexpression further suppressed the invasion and metastasis of CRC. These effects were regulated through the AKT/mTOR signaling pathway.

Molecular mechanisms of azole resistance in Candida tropicalis isolates causing invasive candidiasis in China.

OBJECTIVE: We investigated molecular mechanisms responsible for azole resistance in Candida tropicalis isolates. METHODS: We studied 507 C. tropicalis isolates causing invasive candidiasis from ten hospitals over 5 years. Antifungal susceptibility was determined by broth microdilution methods. Point mutations in the C. tropicalis ERG11 gene that may confer azole resistance were explored and verified. The expression levels of ERG11, CYTb, MDR1 and CDR1 genes were compared in 20 fluconazole-susceptible and 20 fluconazole-resistant isolates. RESULTS: Fluconazole-susceptible, -susceptible dose-dependent and -resistant strains accounted for 76.7% (389/507), 10.5% (53/507) and 12.8% (65/507) of C. tropicalis isolates, respectively. The ERG11 mutation A395T/W occurred in 10.7% (54/507) of isolates, all of which were resistant to fluconazole. The nucleotide mutation C461T/Y was the second most common (50/507 isolates, 9.9%), and all isolates carrying C461T/Y also had the mutation A395T/W. However, the presence of C461T did not contribute to the azole-resistant phenotype. Substitutions V125A, Y257H and G464S (<2% of isolates), which were reported for the first time in C. tropicalis, also conferred fluconazole non-susceptible phenotypes. Compared with fluconazole susceptible isolates, fluconazole-resistant isolates had higher ERG11 (fold expression level 1.42 versus 0.79, p < 0.01) but lower CYTb (fold expression level 1.26 versus 2.67, p < 0.01) gene expression levels. Three azole-resistant isolates carrying the wild-type ERG11 gene had higher levels of CDR1 and MDR1 expression. CONCLUSIONS: ERG11 missense mutations were the major mechanism responsible for azole resistance in C. tropicalis isolates, but overexpression of ERG11, CDR1 and MDR1, as well as reduced expression of CYTb, also contributed to resistance.

Expression of lamin A/C protein in degenerated human intervertebral disc.

OBJECTIVE: This study aimed to evaluate the expression characteristics of lamin A/C proteins in intervertebral disc degeneration (IVD) specimens from patients with different degeneration grades. Lamin A/C proteins have been shown to result in age-related changes in the osteoarticular system. However, the expression characteristics of these nuclear proteins in degenerated human IVD tissues have not been explored previously. PATIENTS AND METHODS: Degenerated human IVD tissues were obtained during spinal surgery. Articular cartilage samples after total knee replacement surgery were used as controls. Sections of these tissues were stained with hematoxylin and eosin, Masson, safranin O, and immunostained using lamin A/C antibody. Western blot was performed to evaluate lamin A/C expression in IVD tissues. Lamin A/C expression was analyzed based on different degeneration grades. RESULTS: In patients with IVD degeneration, mild or moderate degenerative discs contained high amounts of lamin A/C proteins. Lamin A/C expression was primarily localized in the nuclear envelope of IVD cells, and associated with apoptosis in cell nuclei, as determined by immunostaining and TUNEL assay. CONCLUSIONS: This paper is the first to report that lamin A/C proteins are present in IVD tissues and its expression may be related to disc degeneration.

[The role of Th1/Th2 cells imbalance in the pathogenesis of secretory otitis media].

Objective:The aim of this study is to explore the role of Th1/Th2 cells imbalance in the pathogenesis of secretory otitis media. Method:Ninety secretory otitis media patients were enrolled in observation group. According to medical history, they were divided into acute and chronic group. In addition, 90 healthy volunteers during the same period were selected as the control group. The levels of Th1-type cytokines IFN-γ, Th2-type cytokines interleukin-4 (IL-4) and IFN-γ/IL-4 in peripheral blood were compared between observation group and control group. Compare with acute and chronic secretory otitis media patients IFN-γ, IL-4, IFN-γ/IL-4 levels as well as the compare with middle ear effusion and peripheral blood sIFN-γ, IL-4, IFN-γ/IL-4 levels in observation group. Result:The level of IFN-γ, IL-4 and IFN-γ/IL-4 in the peripheral blood in the observation group were higher than those of the control group (P<0.05). The levels of IFN-γ, IL-4 and IFN-γ/IL-4 in the peripheral blood of patients in the chronic group were higher than those in the acute group. There was no significant difference in IL4, IFN-γ/IL-4 levels between the observation group and the middle ear effusion (P>0.05), IFN-γ levels in peripheral blood were lower than those of the middle ear effusion IFN-γ (P<0.05). Conclusion:Abnormal IFN-γ, IL-4 levels of the peripheral blood and the middle ear effusion have some relationship with secretory otitis media, and Th1/Th2 imbalance may be a risk factor for secretory otitis media.

[Expert consensus on the diagnosis and treatment of cryptococcal meningitis].

Cryptococcal meningitis is a common and refractory central nervous system infection, with high rates of mortality and disability. The experts of the Society of Infectious Diseases of Chinese Medical Association have reached this consensus after a thorough discussion. Based on the current situation of cryptococcal meningitis in China, the management of cryptococcal meningitis includes 6 aspects: introduction, microorganism identification, clinical manifestations and diagnosis, principles of antifungal therapy, treatment of refractory and recurrent meningitis, treatment of intracranial hypertension. There is not a separate consensus on human immunodeficiency virus (HIV) infection in patients with cryptococcal meningitis. This article focuses on different antifungal regimens and reducing intracranial pressure by reference to Infectious Disease Society of America (IDSA) guidelines. The importance of early diagnosis, combined long-term antifungal therapy, control of intracranial hypertension are emphasized.

[The composition and antimicrobial resistance of isolates from lower respiratory tract and blood in hospitalized patients in respiratory ward: a multicenter national study in China].

Objective: To investigate the species and antimicrobial resistance of bacterial pathogens isolated from hospitalized patients in respiratory ward in China. Methods: This was a multicenter retrospective study based on a national epidemiological network called China Antimicrobial Resistance Surveillance System (CARSS). The non-repetitive strains isolated from lower respiratory tract and blood samples in 91 hospitals from seven geographic regions of CARSS were reviewed. The distribution of specimen type, hospital level (secondary and tertiary hospital), patient age group [geriatric (>65 years old), adult (15 to 65 years old), pediatric (28 days to 14 years old ) and newborn group (≤28 days)] and ward type (respiratory intensive care unit and general respiratory ward) were analyzed for MRSA, PRSP, CREC, CRKP, CRPA, CRAB, ESBL-EC and ESBL-KP. The categorical variables were analyzed by chi-square test using SPSS 16.0 statistical software. P<0.05 was regarded as statistically significant. Results: A total of 50 417 non-repetitive isolates [42 751 isolates from lower respiratory tract (LRT), 2 649 isolates from blood and 5 017 isolates from other samples (urine and secretions)] from 48 752 inpatients (without illness type information) were enrolled in the study. 90.2% (45 491/50 417) isolates were obtained from 63 tertiary hospitals. According to patients' age, all cases were divided into 4 groups, i. e. geriatric(46.0%, 23 177/50 417), adult(29.9%, 15 092/50 417), pediatric(24.0%, 12 112/50 417) and newborn group(0.0%, 36/50 417). All isolates were obtained from respiratory intensive care unit (6.2%, 3 129/50 417) or general respiratory wards (93.8%, 47 288/50 417). The majority of bacterial pathogens were isolated from lower respiratory and blood culture samples, which accounted for 90.0% of all the samples (45 400/50 417). Sputum accounted for 81.6% (41 131/50 417) of samples, and the leading 4 isolates were K. pneumonia (18.9%, 7 784/41 131), P. aeruginosa (13.6%, 5 580/41 131), A. baumanni (11.3%, 4 644/41 131) and S. pneumonia (11.1%, 4 564/41 131). Blood samples accounted for 5.3% (2 649/50 417) of the samples, with the leading 4 bacteria being coagulase-negative staphylococcus (42.0%, 1 112/2 649), E. coli (18.3%, 484/2 649), K. pneumonia (7.4%, 194/2 649) and S. aureus (4.9%, 131/2 649). The species distribution of pathogens isolated from bronchoalveolar lavage fluid (BALF), which accounted for 3.2% (1 620/50 417) of the samples, was similar to that of sputum, and the leading 4 bacteria were P. aeruginosa (22.0%, 360/1 620), K. pneumonia (14.8%, 239/1 620), A. baumanni (11.9%, 193/1 620) and S. pneumonia (9.6%, 155/1 620). The prevalence of CRKP, CRPA and CRAB in tertiary hospitals [5.2% (384/7 439), 23.8% (1 260/5 304) and 53.5% (2 259/4 224), respectively] was significantly higher than that in secondary hospitals [2.5% (24/973), 12.8% (101/787) and 33.9% (109/322), respectively] (all P<0.01). In comparison, the prevalence of ESBL-EC in secondary hospitals (63.9%, 145/227) was higher than that in tertiary ones (55.0%, 1 141/2 074) (P=0.011). The prevalence of ESBL-EC and ESBL-KP in pediatric group [68.2% (283/415) and 55.3% (183/331), respectively] was higher than that in geriatric group [54.2% (684/1 263) and 27.1% (625/2 303), respectively] and adult group [51.1% (317/620) and 15.1% (272/1 804), respectively] (all P<0.001). Conclusions: In China, the predominant bacterial pathogens in the respiratory wards were Enterobacteriaceae and non-fermentative bacteria. High prevalence of ESBL-EC and ESBL-KP isolated from lower repiratory tract was revealed in primary hospitals and pediatric patients.